# Multi-tissue transcriptomic profiling reveals the internal physiological landscape of laying hens in cage and cage-free systems

**Authors:** Nonoko N. Shimura, Eiki Asagi, Tadahiro Matsubara, Itsufumi Sato, Yuki Higashiura, Saki Nakamura, Chihiro Kase, Atushi J. Nagano, Shozo Tomonaga, Jun-ichi Shiraishi, Kaito Kurogi, Ryohei Matsuo, Shinobu Yasuo, Tatsuhiko Goto, Kan Sato, Tsuyoshi Shimmura

PMC · DOI: 10.1016/j.psj.2026.106650 · 2026-02-18

## TL;DR

This study compares the physiological effects of different housing systems on laying hens using transcriptomic and behavioral data.

## Contribution

The study reveals how cage-free systems affect metabolic and neurophysiological pathways in laying hens.

## Key findings

- Cage-free systems promote higher insulin sensitivity and norepinephrine signaling compared to caged systems.
- Transcriptomic analysis showed insulin resistance in caged hens and enhanced signaling in cage-free hens.
- Behavioral diversity increased in barn and free-range systems compared to battery cages.

## Abstract

Welfare-friendly housing systems for laying hens, such as cage-free, have become prevalent. However, the physiological effects of housing systems on the laying hens remain poorly understood. Here, we compared behavioral characteristics and transcriptomic profiles from 90 multi-tissue samples among three housing systems: battery cage (BC), barn (BR), and free-range (FR). BR and FR housing promoted behavioral diversity compared to cages. Transcriptome analysis of central tissues (diencephalon and cerebral hemisphere) and peripheral tissues involved in egg production (liver, ovary, oviductal segments of magnum, and uterine) revealed significant enrichment of insulin resistance-related pathways in both diencephalon and liver of BC hens, and enhanced norepinephrine signaling in the cerebrum of BR and FR hens. To validate these findings, we performed a glucose tolerance test to assess insulin sensitivity and quantified the cerebral norepinephrine concentrations by ECD-HPLC. The results showed that BR and FR hens tended to exhibit higher insulin sensitivity and enhanced norepinephrine signaling compared with BC hens. Taken together, our findings suggest that housing conditions markedly shape the internal physiological landscape of laying hens, and also that environment-enriched cage-free contributes to improving metabolic and neurophysiological signaling.

## Linked entities

- **Chemicals:** norepinephrine (PubChem CID 951)
- **Species:** Gallus gallus (taxon 9031)

## Full-text entities

- **Genes:** ADRA1D (adrenoceptor alpha 1D) [NCBI Gene 422933], CSF3 (colony stimulating factor 3) [NCBI Gene 396216], CCR10 (C-C motif chemokine receptor 10) [NCBI Gene 100858200], PNMT (phenylethanolamine N-methyltransferase) [NCBI Gene 100858738], CLDN18 (claudin 18) [NCBI Gene 429135], BRAF (B-Raf proto-oncogene, serine/threonine kinase) [NCBI Gene 396239] {aka C-RMIL, RMIL}, ADRA2A (adrenoceptor alpha 2A) [NCBI Gene 428980], COMT (catechol-O-methyltransferase) [NCBI Gene 416783], STAT3 (signal transducer and activator of transcription 3) [NCBI Gene 420027], RPS6KA1 (ribosomal protein S6 kinase A1) [NCBI Gene 386579] {aka RPS6KA, RPS6KA1L, RSK}, CNTN3 (contactin 3) [NCBI Gene 416101] {aka contactin-3}, RPS6KA3 (ribosomal protein S6 kinase A3) [NCBI Gene 418605], ADRB1 (adrenoceptor beta 1) [NCBI Gene 101750705], GLUL (glutamate-ammonia ligase) [NCBI Gene 396489] {aka p42}, LRRC18 (leucine rich repeat containing 18) [NCBI Gene 101751830], CD86 (CD86 molecule) [NCBI Gene 427944], CCL20 (C-C motif chemokine ligand 20) [NCBI Gene 395082] {aka MIP-3ALPHA, Mip-3a, SCYA20, chCCL20}, ATP6V0A1 (ATPase H+ transporting V0 subunit a1) [NCBI Gene 395474] {aka A1}, LOC100859819 (cocaine- and amphetamine-regulated transcript protein-like) [NCBI Gene 100859819] {aka CARTL}, AGRP (agouti related neuropeptide) [NCBI Gene 427535], CARTPT (CART prepropeptide) [NCBI Gene 100858488] {aka CART}, HTR7L (5-hydroxytryptamine receptor 7 like) [NCBI Gene 422942] {aka HTR7C}, ADRB3 (adrenoceptor beta 3) [NCBI Gene 430991], HTR2B (5-hydroxytryptamine receptor 2B) [NCBI Gene 395581], VAT1 (vesicle amine transport 1) [NCBI Gene 420009], MLXIPL (MLX interacting protein like) [NCBI Gene 101751014] {aka ChREBP}, PCSK2 (proprotein convertase subtilisin/kexin type 2) [NCBI Gene 395136] {aka PC2}, CDH1 (cadherin 1) [NCBI Gene 415860] {aka cadherin-1}, OVAL (ovalbumin (SERPINB14)) [NCBI Gene 396058] {aka OVA, SERPINB14}, LEP (leptin) [NCBI Gene 373955] {aka OB}, PCK1 (phosphoenolpyruvate carboxykinase 1) [NCBI Gene 396458] {aka PEPCK}, INS (insulin) [NCBI Gene 396145], IRX3 [NCBI Gene 408181], CLDN3 (claudin 3) [NCBI Gene 374029] {aka claudin-3}, PIGR (polymeric immunoglobulin receptor) [NCBI Gene 419848], RPS6KA2 (ribosomal protein S6 kinase A2) [NCBI Gene 421570], CFAP77 (cilia and flagella associated protein 77) [NCBI Gene 417168] {aka C17H9ORF171}, CATH2 (cathelicidin 2) [NCBI Gene 420407] {aka CAMP, CATH-2, CATHL2, CMAP27}, MYLK (myosin light chain kinase) [NCBI Gene 396445] {aka KRP, MLCK}, SLC18A1 (solute carrier family 18 member A1) [NCBI Gene 431330], AKT3 (AKT serine/threonine kinase 3) [NCBI Gene 421497]
- **Diseases:** chronic (MESH:D002908), Liver (MESH:D017093), depression (MESH:D003866), foot damage (MESH:D018409), Insulin resistance (MESH:D007333), fatty liver hemorrhagic syndrome (MESH:D005234), gain (MESH:D015430), anxiety (MESH:D001007), wounds (MESH:D014947), inflammation (MESH:D007249)
- **Chemicals:** 5-HT (MESH:D012701), Glucose (MESH:D005947), calcium (MESH:D002118), acetate (MESH:D000085), sodium acetate (MESH:D019346), DA (MESH:D004298), perchloric acid (MESH:C576518), lipid (MESH:D008055), citrate (MESH:D019343), 3-Methoxy-4-hydroxy-phenylglycolaldehyde (-), 5-HIAA (MESH:D006897), blood glucose (MESH:D001786), NE (MESH:D009638), E (MESH:D004540), IP3 (MESH:D015544), tyrosine (MESH:D014443), water (MESH:D014867), 3,4-Dihydroxyphenylacetic acid (MESH:D015102), triglyceride (MESH:D014280), Epinephrine (MESH:D004837), phosphorus (MESH:D010758), methanol (MESH:D000432)
- **Species:** Gallus gallus (bantam, species) [taxon 9031], Homo sapiens (human, species) [taxon 9606]

## Figures

5 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12974103/full.md

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Source: https://tomesphere.com/paper/PMC12974103